Stretch Slows Conduction in the Human Atrium. Introduction: The mechanisms by which atrial stretch favors the development of a substrate for atrial fibrillation (AF) are not fully understood. In this study, the role of stretch-induced conduction changes in the creation of a proarrhythmic substrate has been investigated by quantifying the spatial distribution of local conduction velocities (CVs) in the human atrium during acute atrial dilatation. Methods and Results: Electroanatomic mapping of right atrial activation was performed in 10 patients during coronary sinus pacing under control condition and during acute atrial dilatation. Atrial stretch was obtained by simultaneous atrioventricular (AV) pacing at a cycle length of 450-500 ms. Local CVs were accurately estimated by applying the principle of triangulation and spatially mapped over the whole right atrial endocardial surface. Simultaneous AV pacing significantly increased right atrial volume from 72.0 ± 29.0 to 86.3 ± 31.3 mL (P < 0.001). The 23% increase in atrial volume resulted in an overall decrease in atrial CV from 65.8 ± 5.9 to 55.2 ± 7.2 cm/s (P < 0.001) and an increased incidence of slow conduction sites or local conduction blocks from 10.3 ± 4.2% to 15.9 ± 7.7% (P < 0.01). Acute atrial dilatation concurrently increased AF vulnerability, with 6 of 10 patients developing AF episodes under stretch condition. Conclusion: Quantification of stretch-induced conduction changes in the human atrium is feasible by combining simultaneous AV pacing and CV map construction. Acute atrial dilatation results in conduction slowing and significant increase in AF vulnerability, suggesting the role of stretch-induced conduction disturbances in the creation of a substrate for AF. (J Cardiovasc Electrophysiol, Vol. 22, pp. 394-401).